GENETIC REGULATION OF ALCOHOL METABOLISM IN YEAST

目的是了解真核基因的调控 在核酸-蛋白质相互作用和代谢方面， 调节效应分子。 具体目标是 表征调控表达所需的DNA序列 并鉴定和表征与之相互作用的蛋白质 这些序列。 22个核苷酸的特殊变化 碱基对二联体（上游激活位点，或UAS）， 调节酵母醇脱氢酶2基因 将被构造和表征，以确定潜在点 ADR 1是一种蛋白质， 在无人机系统中相互作用 ADR 1介导的结构域 核进入，DNA结合，转录激活，和 将通过分析缺失突变体来鉴定调控。 的 推定的DNA结合和转录激活结构域 将通过经典和重组遗传学分析ADR 1 （使用寡核苷酸定向诱变）和通过构建 杂合调控基因 一个假定的TATA或 由ADR 6编码的CAP调节因子将通过以下方式实现： 遗传和生化手段。 更好地了解酒精代谢和基因 控制乙醇的合成和降解， 这些研究的医疗效益。 基本了解如何 控制基因作用的蛋白质最终将对医学有益 也

The objective is to understand the regulation of a eukaryotic gene in terms of nucleic acid-protein interaction and metabolic regulation of effector molecules. The specific aims are to characterize the DNA sequences required for regulated expression and to identify and characterize the proteins which interact with these sequences. Specific nucleotide changes in the twenty-two base pair dyad (the upstream activation site, or UAS) which mediates regulation of the alcohol dehydrogenase 2 gene of yeast will be constructed and characterized to identify potential points of contact between the DNA and ADR1, the protein which apparently interacts at the UAS. The domains of ADR1 mediating nuclear entry, DNA binding, transcriptional activation, and regulation will be identified by analysis of deletion mutants. The putative DNA binding and transcription activation domain of ADR1 will be analyzed by classical and recombinant genetics (using oligonucleotide directed mutagenesis) and by constructing hybrid regulatory genes. Characterization of a putative TATA or CAP regulator factor encoded by ADR6 will be achieved by genetic and biochemical means. A better understanding of alcohol metabolism and the genes controlling ethanol synthesis and degradation are anticipated medical benefits of these studies. A basic understanding of how proteins control gene action will ultimately be of medical benefit as well.